Rapid nodal staging of head and neck cancer surgical specimens with flow cytometric analysis

Häyry, Valtteri; Kågedal, Åsa; Hjalmarsson, Eric; Silva, Pedro Farrajota Neves da; Drakskog, Cecilia; Margolin, Gregori; Georén, Susanna Kumlien; Munck‐Wikland, Eva; Winqvist, Ola; Cardell, Lars Olaf

doi:10.1038/bjc.2017.408

Cited by 14 publications

(9 citation statements)

References 17 publications

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“…Similarly, Leers and colleagues have shown that multiparameter flow cytometry is useful for the detection of micrometastasis in SLN of breast cancer patients [30] and Hartana et al in renal tumors [31]. More recently, Häyry and colleagues have shown that rapid nodal staging of head and neck cancer could be achieved with flow cytometry, demonstrating the fast and inexpensive nature of this new perioperative staging method [32]. Jagric and colleagues have demonstrated that for high risk SLN from gastric cancer patients, flow cytometry is highly specific in cancer detection although should not be used as a stand-alone method intra-operatively due to low sensitivity [33].…”

Section: Discussionmentioning

confidence: 96%

Cytomorphological Characterization of Individual Metastatic Tumor Cells from Gastrointestinal Cancer Patient Lymph Nodes with Imaging Flow Cytometry

Winter

Gibson

Ruszkiewicz

et al. 2019

Gastrointestinal Disorders

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The presence or absence of tumor cells within patient lymph nodes is an important prognostic indicator in a number of cancer types and an essential element of the staging process. However, patients with the same pathological stage will not necessarily have the same outcome. Therefore, additional factors may aid in identifying patients at a greater risk of developing metastasis. In this proof of principle study, initially, spiked tumor cells in rat lymph nodes were used to mimic a node with a small cancer deposit. Next, human lymph nodes were obtained from cancer patients for morphological characterization. Nodes were dissociated with a manual tissue homogenizer and stained with fluorescent antibodies against CD45 and Pan-Cytokeratin and then imaging flow cytometry (AMNIS ImageStreamX Mark II) was performed. We show here that imaging flow cytometry can be used for the detection and characterization of small numbers of cancer cells in lymph nodes and we also demonstrate the phenotypical and morphological characterization of cancer cells in gastrointestinal cancer patient lymph nodes. When used in addition to conventional histological techniques, this high throughput detection of tumor cells in lymph nodes may offer additional information assisting in the staging process with therapeutic and prognostic applications.

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Section: Discussionmentioning

confidence: 96%

Cytomorphological Characterization of Individual Metastatic Tumor Cells from Gastrointestinal Cancer Patient Lymph Nodes with Imaging Flow Cytometry

Winter

Gibson

Ruszkiewicz

et al. 2019

Gastrointestinal Disorders

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“…We hypothesized that sentinel node-derived T cells are a more reliable and representative source for evaluation of affected patient’s oncoimmunological status and therefore have potential to provide better insight into their prognosis and choice of treatment. Taking into account our experience in flow cytometry analysis of TDLN 14 , we prospectively gathered a cohort of OSCC patients and analyzed TILs, T-cells activation in primary tumour, sentinel, non-sentinel nodes and PBMC in relation to recurrence status. Influence of the aforementioned variables on survival was studied to identify prognostic markers.…”

Section: Introductionmentioning

confidence: 99%

Activation of T helper cells in sentinel node predicts poor prognosis in oral squamous cell carcinoma

Kågedal

Hjalmarsson

Silva

et al. 2020

Sci Rep

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Recurrence in oral squamous cell carcinoma (OSCC) significantly reduces overall survival. Improved understanding of the host’s immune status in head and neck cancer may facilitate identification of patients at higher risk of recurrence and improve patients’ selection for ongoing clinical trials assessing the effectiveness of immune checkpoint inhibitors (CPI). We aimed to investigate Sentinel Node-derived T-cells and their impact on survival. We enrolled prospectively 28 OSCC patients treated at Karolinska University Hospital, Stockholm, Sweden with primary tumour excision and elective neck dissection. On top of the standard treatment, the enrolled patients underwent sentinel node procedure. T cells derived from Sentinel nodes, non-sentinel nodes, primary tumour and PBMC were analyzed in flow cytometry. Patients who developed recurrence proved to have significantly lower level of CD4+ CD69+ in their sentinel node (31.38 ± 6.019% vs. 43.44 ± 15.33%, p = 0.0103) and significantly higher level of CD8+ CD HLA-DR+ (38.95 ± 9.479% vs. 24.58 ± 11.36%, p = 0.0116) compared to disease-free individuals. Survival analysis of studied population revealed that patients with low proportion of CD4+ CD69+ had significantly decreased disease-free survival (DFS) of 19.7 months (95% CI 12.6–26.9) compared with 42.6 months (95% CI 40.1–45.1) in those with high CD4+ CD69+ proportion in their Sentinel Nodes (log-rank test, p = 0.033). Our findings demonstrate that characterization of T-cell activation in Sentinel Node serves as a complementary prognostic marker. Flow cytometry of Sentinel Node may be useful in both patients’ surveillance and selection for ongoing CPI clinical trials in head and neck cancer.

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Section: Introductionmentioning

confidence: 99%

“…Non hemopoietic neoplasms may be identified by FC in tissues looking for the cytoplasmic expression of cytokeratins in case of carcinomas or for the surface expression of CD56 in case of neuroendocrine tumours. [7][8][9][10] FC cytokeratin evaluation however may be not always possible, as cytoplasmic antigen detection implies cell fixation and permeabilization, which can cause further cell loss in paucicellular or bloody specimens, as frequently occurring in FNA and body fluids. The use of directly conjugated antibodies towards CD326 (EpCAM), the epithelial cell adhesion molecule present on the surface and in the cytoplasm of all epithelial cells, 11 allows their rapid identification without extensive sample manipulations.…”

mentioning

confidence: 99%

Flow cytometry identification of nonhemopoietic neoplasms during routine immunophenotyping

Stacchini

Aliberti

Demurtas

et al. 2018

Int J Lab Hematology

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Introduction Nonhemopoietic neoplasms (NHNs) may be encountered during routine flow cytometry (FC) immunophenotyping. The clue of their presence mainly relies on detection of CD45‐negative (CD45−) cells with altered scatter parameters. Methods In this study, we evaluated a monoclonal antibody combination conceived to characterize the CD45− population by FC, suspected of belonging to NHNs, when present. The panel included CD45 for leucocytes identification, CD326 (clones BerEP4 and HEA‐125) to mark epithelial cells, CD33 to identify myeloid cells, CD138 to trace plasma cells and CD56 useful in the identification of neuroendocrine tumours. 7AAD vital dye was used to gate out dead cells. Results were correlated with cytomorphology and confirmed by histological data, if available. Results Among 9422 specimens submitted for routine FC investigation, 47 samples that included fine‐needle aspirates, bone marrow aspirates, tissue biopsies and body fluids had a detectable CD45− population and a sufficient cell amount to be further investigated. FC revealed the presence of CD326‐positive epithelial cells in 38 specimens; altered scatter parameters and variable reactivity to the other antigens tested allowed to suspect NHNs in the remaining nine samples. The presence of NHNs was confirmed in all cases by morphology. Conclusions The current results show that when CD45− cells with altered scatter parameters were detected, cytometrists involved in leukaemia/lymphoma diagnosis may require further FC investigations to rapidly identify NHNs in different specimens, thus reducing the time of the immunohistochemical diagnostic workup to reach a final diagnosis.

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Rapid nodal staging of head and neck cancer surgical specimens with flow cytometric analysis

Abstract: We show that flow cytometric analysis of nodal tissue is sensitive and reliable in identifying metastases of OSCC. Flow cytometry is inexpensive and fast, providing a possibility of perioperative diagnostics and immediate treatment planning.

Cited by 14 publications

References 17 publications

Cytomorphological Characterization of Individual Metastatic Tumor Cells from Gastrointestinal Cancer Patient Lymph Nodes with Imaging Flow Cytometry

Cytomorphological Characterization of Individual Metastatic Tumor Cells from Gastrointestinal Cancer Patient Lymph Nodes with Imaging Flow Cytometry

Activation of T helper cells in sentinel node predicts poor prognosis in oral squamous cell carcinoma

Flow cytometry identification of nonhemopoietic neoplasms during routine immunophenotyping

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